目的: 通过分析STMN1的表达水平与放疗局部控制率的相关性,初步探讨STMN1的表达与NSCLC患者预后及近期放疗疗效是否相关。方法: 利用免疫组织化学（immumohistochemical staining,IHC）检测STMN1在55例石蜡包埋NSCLC肿瘤组织中及42例正常肺组织中的表达,分析STMN1表达水平与患者临床病理特征、无进展生存期（progression-free survival,PFS）的相关性,进一步通过单因素、多因素回归分析证实STMN1是否为 Ⅲ — Ⅳ期NSCLC患者放疗抵抗的独立预后因素。慢病毒感染方法构建A549及H1299细胞STMN1过表达质粒及干扰细胞模型,通过蛋白免疫印迹技术（Western-blot,WB）验证STMN1和p-STMN1的表达情况,并经X射线处理后,通过流式细胞凋亡检测分析考察STMN1对NSCLC细胞放疗敏感性影响。Transwell侵袭及迁移实验考察STMN1对细胞侵袭及迁移能力的影响。结果: NSCLC肿瘤组织中STMN1的表达水平显著高于正常肺组织;STMN1的表达水平与NSCLC患者的肿瘤分化程度和TNM分期显著相关,与年龄、性别、组织病理学分型无明显相关性;STMN1高表达NSCLC患者中位PFS显著低于STMN1低表达患者;STMN1高表达NSCLC患者放疗2个月后局部控制率显著低于STMN1低表达患者。STMN1中N端Ser的扩增是保证其磷酸化的关键。STMN1过表达可抑制X射线诱发的肺癌细胞凋亡;下调STMN1表达可促进X射线诱发的肺癌细胞凋亡。STMN1过表达后A549及H1299细胞株侵袭及迁移能力显著增强;下调STMN1表达,细胞株侵袭及迁移能力受到抑制。结论: 与正常肺组织相比,STMN1在NSCLC组织中呈现高表达水平,其高表达水平与肿瘤低分化、TNM分期密切相关,同时STMN1高表达NSCLC患者放疗预后差。体外细胞实验证实上调STMN1表达的NSCLC细胞侵袭及迁移能力提高,经X射线辐射干预后,凋亡减少。因此,STMN1可作为NSCLC患者预后不良及放疗疗效抵抗的指标。
Abstract
Objective To investigate whether the expression of STMN1 is related to the prognosis of non-small cell lung cancer (NSCLC) and the effect of radiotherapy. Methods Immunohistochemical staining (IHC) was used to detect the expression of STMN1 in NSCLC and normal tissues. The correlation between STMN1 expression level and clinicopathological characteristics of patients was analyzed, and the correlation between STMN1 expression level and progression-free survival (PFS) was studied. STMN1 was identified as an independent prognostic factor for radiotherapy resistance in stage III-IV NSCLC by univariate and multivariate regression analysis. Constructing STMN1 overexpressing plasmids and interfering cell models of A549 and H1299 cells by a lentivirus infection method, verifying the expression of STMN1 and p-STMN1 by a protein immunoblot technology (Western-blot, WB), The effect of STMN1 on radiosensitivity of NSCLC cells was investigated by flow cytotoxicity analysis. The effect of STMN1 on cell invasion and migration was investigated by transwel1 invasion and migration assay. Results The expression level of STMN1 in NSCLC was significantly higher than that in normal lung. The expression level of STMN1 was significantly correlated with tumor differentiation and TNM staging in NSCLC patients, but not with age, sex and histopathological classification. The median PFS in STMN1 overexpression NSCLC was significantly lower than that in STMN1 underexpression. The local control rate of STMN1 overexpression NSCLC after 2 months of radiotherapy was significantly lower than that of STMN1 underexpression. Overexpression of STMN1 inhibited X-ray induced apoptosis of lung cancer cells. After STMN1 overexpression, the invasion and migration of A549 and H1299 cells were significantly enhanced. Conclusion The high expression of STMN is closely related to the low differentiation and TNM staging of tumor. Therefore, STMN1 can be used as an indicator of poor prognosis and resistance to radiotherapy in patients with NSCLC.
关键词
STMN1 /
非小细胞肺癌 /
放疗耐受 /
不良预后
Key words
STMN1 /
non-small cell lung cancer /
radiotherapy resistence /
poor prognosis
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基金
湖南省自然科学基金项目“STMN1 Ser25与Ser38位点磷酸化结合GRP78促进肺癌细胞侵袭转移及放疗抵抗”（2021JJ31035）
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